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Don’t Let the Cells Pass You By
Whether you use a high-end instrument or a benchtop cytometer, you'll find helpful tips and the latest research posted by the thousands of scientists who follow the Life Technologies Flow Cytometry page on Facebook.

FEATURED NEW PRODUCTS

View Phagocytosis in Green and Red—pHrodo™ BioParticles® Conjugates

what they are
Proprietary, pH-sensitive pHrodo™ conjugates of E. coli, S. aureus, and zymosan are designed to be specific sensors of phagocytosis. In addition to red-fluorescent versions, we now offer green-fluorescent pHrodo™ BioParticles® conjugates that can be visualized using standard fluorescein filters.

what they offer

Optimized, fixable probes for detecting phagocytosis in live cells

Available as E. coli, S. aureus, and zymosan conjugates of pHrodo™ dye in green and red

how they work
During phagocytosis, cells ingest the pHrodo™ dye-conjugated BioParticles and form phagosomes. These phagosomes fuse with the early lysosome to form the acidic phagolysosome. The nonfluorescent pHrodo™ dye, conjugated to the surface of the BioParticles® reagents, becomes fluorescent with this reduction in pH, making these particles ideal reagents for the study of phagocytosis and its regulation by drugs and environmental factors. Because pHrodo™ dye is minimally fluorescent at neutral pH, wash steps and quencher dyes are not required. To facilitate multiplexing, cells assayed for phagocytic activity with pHrodo™ BioParticles® conjugates can be fixed, preserving the fluorescent signal potentially for up to 24 hours.

Compatibility—use in fluorescence imaging, high content screening (HCS), flow cytometry, high-throughput screening (HTS), and with benchtop instruments such as the Tali®, FLoid®, and Attune® systems

how they work
When incubated with cells, Click-iT® LAA incorporates into cellular membranes. Upon lipid peroxidation, LAA is oxidized and produces 9- and 13-hydroperoxy-octadecadienoic acid (HPODE). These hydroperoxides decompose to multiple α,β-unsaturated aldehydes, which readily modify proteins at nucleophilic side chains. These alkyne-containing modified proteins can subsequently be detected using Click-iT® chemistry and multiplexed with other probes appropriate for fixed cells. Click-iT® LAA reagent can be detected with haptens or with azide-containing fluorophores such as the bright, photostable Alexa Fluor® azide reagents. The Click-iT® Cell Buffer Kit provides researchers with the tools required to perform the click reaction with these reagents.

Detect Retinoblastoma—New Rb ABfinity™ Recombinant Rabbit Antibodies

what they are
Rb, or retinoblastoma, is a tumor suppressor protein in the pocket protein family. A nuclear protein that is widely expressed in retinal cells, pRb prevents the replication of damaged DNA by preventing progression through the cell cycle from G1 to S. pRb is inactivated by phosphorylation and activated by dephosphorylation. Active (dephosphorylated) pRb binds and inactivates the cellular transcription factor E2F1, the function of which is required for cell cycle progression. Aurora B directly phosphorylates Rb at serine 780 both in vitro and in vivo. This novel interaction plays a critical role in regulating the postmitotic checkpoint to prevent endoreduplication after an aberrant mitosis. Phosphorylation of serine 249 and threonine 252 is catalyzed by cyclin D-cdk4. We now offer ABfinity™ recombinant rabbit antibodies specific for Rb.

what they offer

Consistent lot-to-lot performance

Minimize the need to revalidate working antibody dilutions for your experiments each time you order

how they work
ABfinity™ antibodies are manufactured by transfecting mammalian cells with high-level expression vectors containing immunogen-specific heavy- and light-chain antibody cDNA. ABfinity™ oligoclonal antibodies are a mixture of recombinant monoclonal antibodies, combining the improved signal strength of a polyclonal antibody with the highly reproducible results you get from ABfinity™ monoclonal antibodies.

Add, Incubate, and Image—Trial-Size Markers for Cell Cycle and Function

what they are
Our popular Premo™ FUCCI and CellLight® reagents use BacMam technology to provide highly efficient and transient fluorescent protein labeling of live cells, including neurons and stem cells. New small pack sizes make it easy to test these premier live-cell markers with your cells.

what they offer

High efficiency—Greater than 90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons

Speed and convenience—simply add one or more BacMam reagents to your cells, incubate overnight, and image; or store frozen, assay-ready cells for later use

Robustness—expression can be easily adjusted by the dose; no replication in mammalian cells; lack of observable cytopathic effect; biosafety level (BSL) 1 classification

how they work
CellLight® reagents combine the utility and selectivity of fluorescent proteins (CFP, GFP, and TagRFP) with the transduction efficiency of BacMam 2.0, enabling unambiguous staining of organelles and cellular structures in live mammalian cells. Premo™ FUCCI Cell Cycle Sensor is a fluorescent, two-color sensor of cell cycle progression and division in live cells. Cells change from red in the G1 phase, to yellow in G1/S interphase, to green in the S, G2, and M phases. Both reagents are easy to use—simply add, incubate, and image.

Cell cycle imaging with Premo™ FUCCI Cell Cycle Sensor. The cell cycle phases of dividing U2OS cells were visualized for 16 hr using the Premo™ FUCCI Cell Cycle Sensor. The cell in the center of the image transitions from G2/M (green), to G1 (red), and finally to S phase (yellow). After about 6 hr, the cell at the top of the image migrates down before undergoing mitosis (green) and progressing into G1 phase (red).

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Investigate Oxidative Stress and Autophagy in Live Cells

Aberrant generation of reactive oxygen species (ROS) through oxidative stress causes the disruption of normal cell mechanisms, and may lead to cell death. Although the role of autophagy in oxidative stress–induced cell death is not fully understood, the segregation and delivery of cytoplasmic cargo for degradation play a vital role in the cell survival response during early stage, ROS-induced stress. The LC3B protein generally resides in the cytosol, but following cleavage and lipidation with phosphatidylethanolamine, LC3B associates with the autophagosome to sequester oxidized or dysfunctional intracellular components for downstream degradation. This autophagosome ultimately fuses with the lysosome to form a structure known as the autolysosome for final degradation of damaged cytosolic materials. Life Technologies offers a wide variety of reagents for the investigation of oxidative stress and autophagy in live-cell labeling and detection experimentation.

Studies in a variety of different model organisms have shown that new protein synthesis is required for long-term synaptic changes and the formation of long-term memory. Modified amino acids, such as our Click-iT® AHA (L-azidohomoalanine) and Click-iT® HPG (L-homopropargylglycine) can be incorporated into proteins during active synthesis and labeled in a highly specific azide–alkyne click reaction to identify nascent proteins without using radioactivity. This technique has previously been used to examine new protein synthesis in vitro. However, Schuman and colleagues have recently labeled newly synthesized proteins in live zebrafish using Click-iT® AHA. After in vivo incorporation of Click-iT® AHA, zebrafish lysates were labeled with biotin alkyne, and biotinylated proteins were quantified and detected using dot blots. Using an Alexa Fluor® 488 alkyne, the authors were also able to image a whole-mount zebrafish to localize areas of new protein synthesis within the intact organism. The team further showed that in vivo labeling of nascent proteins with Click-iT® AHA did not affect several zebrafish behaviors and that induction of seizures results in an increase in protein synthesis that can be detected by Click-iT® AHA and the click labeling reagents. Based on their findings, the Click-iT® AHA and HPG amino acids could prove useful in examining new protein synthesis that occurs in live animals during memory formation as well as determining the specific proteins that are involved in long-term synaptic changes underlying these behaviors.

The Molecular Probes® Educational Webinar Series

Molecular Probes® scientists have been providing first-in-class support for our innovative fluorescent reagents and labeling technologies for years. To share this expertise with more researchers, we have a series of free webinars focused on key fluorescence applications. Choose your learning module and get started today.